Difference between revisions of "Part:BBa K633000"

Revision as of 00:57, 19 October 2011

CelD Mutated Cellulase

Clostridium thermocellum endoglucanase CelD was chosen as a representative member for detailedstructural and functional studies of the family E cellulases. Beside C. thermocellum’s CelD, E family cellulases include a number of cellulases of bacterial, fungal, and plant origin.
CelD, has been previously overexpressed in Escherichia coli, the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies. (Chauvaux, Beguin, Aubert, 1992)
The CelD sequence used has an aminoacid substittition Asp-523 ---> Ala. That mutation increases the specific activity of the enzyme by 224% (Chauvaux, Beguin, Aubert, 1992)

Chauvaux, S., Beguin, P., & Aubert, J. (1992). Site-directed mutagenesis of essential carboxylic residues in clostridium thermocellum endoglucanase celd*. The Journal of Biological Chemistry, 267(5), 4472-4478. Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1831
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 644
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 508
Illegal AgeI site found at 151
Illegal AgeI site found at 567
Illegal AgeI site found at 1396
1000
COMPATIBLE WITH RFC[1000]

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 <br>
 <br>
-Clostridium thermocellum endoglucanase CelD was chosen as a representative member for detailed structural and functional studies of family E cellulases. Family E includes, beside C. thermocellum CelD, a number of cellulases of bacterial, fungal, and plant origin.
+CelD Mutated Cellulase
-<br>
-<br>
-The corresponding gene, celD, has been overexpressed in Escherichia coli, and the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies.
+Clostridium thermocellum endoglucanase CelD was chosen as a representative member for detailedstructural and functional studies of the family E cellulases. Beside C. thermocellum’s CelD, E family cellulases include a number of cellulases of bacterial, fungal, and plant origin.
-<br>
 <br>
-This specific cellulase sequence is the result of an investigation regarding activity, several mutations directed at the carboxilic residues (aspartate, glutamate), which are involved in the catalytic mechanism of celD, finally obtaining a celD with a 224% specific activity caused by the mutation Asp-523 ---> Ala on the peptide sequence.
+CelD, has been previously overexpressed in Escherichia coli, the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies. (Chauvaux, Beguin, Aubert, 1992)
 <br>
+The CelD sequence used has an aminoacid substittition Asp-523 ---> Ala. That mutation increases the specific activity of the enzyme by 224% (Chauvaux, Beguin, Aubert, 1992)
 <br>
 [[Image:Celd_chart.png]]__NOTOC__